The ProcartaPlex Mouse Immuno-Oncology Checkpoint Panel 4plex enables cancer research by analyzing 4 protein targets: BTLA, CD27, LAG-3, and TIM-3 in a single well using Luminex xMAP technology. This panel allows simultaneous detection of the soluble forms of proteins that play a crucial role in the regulation of T cells, leading to either T cell exhaustion or stimulation, thus modifying the antitumor immune response. Analysis of these soluble biomarkers could help shed light on the biology of the pathways. The panel is provided in a ready-to-use format with individual vials of capture and detection reagents formulated at 1X concentration, requiring less pipetting and experimental setup. It is not combinable with simplexes or other panels. It complements the Mouse Immuno-Oncology Checkpoint Panel 2 7plex (Cat. No. EPX070-20835-901) that provides 7 additional protein targets in the same research area.

ProcartaPlex Preconfigured panels are extensively tested for analyte combinability, interference, and cross-reactivity to provide the highest level of validation and precision. All ProcartaPlex panels are supplied with the necessary reagents to perform the assay.

Target list [bead region]: BTLA [61], CD27 [75], LAG-3 [44], TIM-3 [46]

About ProcartaPlex assays for the Luminex platform

ProcartaPlex immunoassays are based on the principles of a sandwich ELISA, using two highly specific antibodies binding to different epitopes of one protein to quantitate all protein targets simultaneously using a Luminex instrument. ProcartaPlex multiplex assays require as little as 25 μL of plasma or serum, or 50 μL of cell culture supernatant, and just four hours to obtain analyzed results.

• Reproducible, reliable results—validated as a panel to the highest industry standard, including protein target combinability and cross-reactivity testing
• More results per sample—measure multiple protein targets simultaneously in a single 25–50 μL sample
• Well-established Luminex technology—highly referenced multiplexing platform for protein detection and quantitation

ProcartaPlex assays utilize Luminex xMAP (multianalyte profiling) technology for the simultaneous detection and quantitation of up to 80 protein targets in a single 25–50 μL sample—from plasma, serum, cell culture supernatants, and other bodily fluids.

The Luminex beads in the ProcartaPlex assay are internally dyed with precise proportions of red and infrared fluorophores to create spectrally unique signatures that can be identified by the Luminex xMAP detection systems (e.g., Luminex 200, FLEXMAP 3D, and MAGPIX). Similar to a sandwich ELISA, the ProcartaPlex assay uses matched antibody pairs to identify the protein of interest. In a multiplexed assay, each spectrally unique bead is labeled with antibodies specific for a single target protein, and bound proteins are identified with biotinylated antibodies and streptavidin–R-phycoerythrin (RPE). The conjugation of protein-specific antibodies to a distinct bead allows for analysis of multiple targets in a single well.

The most significant difference between a ProcartaPlex assay and ELISA is that the capture antibody in the ProcartaPlex assay is conjugated to a bead and not adsorbed to the microplate well, so the ProcartaPlex assay reagents are free-floating in the solution.